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用于活细胞成像的超分辨光学显微镜方法与应用
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Abstract:
细胞是生命体活动的基本单元，对活细胞的实时观测有助于在更接近生理的条件下观察其细微结构及其动力学过程，理解生命的本质。近三十年，超分辨光学显微成像技术(Super-Resolution Microscopy, SRM)和相关技术的发展，允许人们在突破衍射极限的尺度下对活细胞进行观察与研究，然而这些技术早期应用于活细胞成像领域时遭受到了不同程度的挑战，而随着后续荧光染料等相关技术的发展，SRM在活细胞成像领域的应用愈加广泛。本文通过简要介绍目前常见的几种SRM以及近些年受到广泛关注的最小光子通量(MINFLUX)等显微技术的基本原理和特点，梳理了其在活细胞成像领域的最新应用。
As the fundamental unit of life activities, the real-time observation of live cells facilitates the investigation of their intricate structures and dynamic processes under conditions closer to physiological states, thereby advancing our understanding of life’s essence. Over the past three decades, advancements in super-resolution microscopy (SRM) and related technologies have enabled the observation and study of live cells at scales surpassing the diffraction limit. However, the initial application of these techniques in live-cell imaging faced significant challenges. With subsequent advancements in fluorescent dyes and other ancillary technologies, SRM has become increasingly applied in live-cell imaging. This review provides a concise introduction to the principles and characteristics of several widely used SRM techniques, as well as emerging methodologies such as minimal photon flux (MINFLUX) microscopy, which has garnered substantial attention in recent years. Additionally, we summarize their latest applications and breakthroughs in the field of live-cell imaging.
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